Filoviruses, such as Ebola virus (EBOV), are CDC category A agents that cause severe hemorrhagic fever syndromes. Unfortunately, few therapeutic options are available to treat infections with these viruses. Apath has an ongoing screening program that utilizes a cell-based, infection-independent assay based on an EBOV minigenome-expression system. The goal of this proposal is to advance the development of EBOV inhibitors that we have identified in this proprietary screening assay. We have confirmed anti-Ebola virus activity for several compounds in viral assays performed at USAMRIID and testing of others is ongoing. For this Bioshield initiative we propose to bring these potential anti-EBOV lead compounds through a preclinical program of absorption, distribution, metabolism, elimination (ADME), toxicology (Tox), and pharmacokinetic (PK) studies in preparation for an eventual investigational new drug (IND) application to the FDA. These studies will be performed in two stages. Initially a preliminary evaluation will be performed to predict how the antiviral compounds will eventually behave in studies performed with full regulatory protocols. These preliminary analyses will include cytotoxicity, solubility, metabolic stability, cell permeability, plasma protein binding, and CYP inhibition. The compounds that demonstrate the most promising profiles in these preliminary analyses will be advanced to a full program of ADME-Tox and PK studies performed under good laboratory practices (GLP) regulations to obtain data to support an IND application. It is expected that these studies will result in the production of antiviral drugs for future clinical trials to determine safety and efficacy against EBOV infection.